BACKGROUND:SB216763 has demonstrated efficacy in treating pulmonary fibrosis in animal models, but its underlying mechanisms remain poorly understood. This study used network pharmacology, molecular docking, and molecular dynamics simulations to explore the molecular mechanisms by which SB216763 exerts therapeutic effects on pulmonary fibrosis. METHODS:The targets of SB216763 were screened using the PharmMapper database, while pulmonary fibrosis-related targets were obtained from GeneCards and OMIM databases. A Venn diagram generated from these datasets highlighted overlapping targets. Protein-protein interaction networks were constructed using the STRING database and Cytoscape software to assess target importance. GO and KEGG enrichment analyses using the Metascape database revealed relevant biological functions and signaling pathways of SB216763 in the treatment of pulmonary fibrosis. Molecular docking investigated the binding affinity and binding mode of SB216763 with GSK-3β. Molecular dynamics simulations further confirmed the stability of the complex formed by SB216763 and GSK-3β. Finally, Western blotting, Griess assays, and ELISA were employed to validate the effect of SB216763 on macrophage functional plasticity in pulmonary fibrosis. RESULTS:Network pharmacology analysis identified the following three key targets of SB216763 in pulmonary fibrosis: AKT1, GSK3B, and TGFBR1, along with the PI3K-AKT signaling pathway. Molecular docking and dynamics simulations revealed that SB216763 primarily interacts with GSK-3β through hydrogen bonds and hydrophobic interactions. In vitro validation demonstrated significant anti-inflammatory and antifibrotic effects of SB216763. Additionally, SB216763 exhibited good safety in macrophages. CONCLUSIONS:This study provided new insights into the mechanisms of SB216763 in the treatment of pulmonary fibrosis.
The recent decision to establish a single pivotal trial as the default evidentiary standard for Food and Drug Administration (FDA) approval marks a substantive shift in the architecture of regulatory proof. This marks a shift from replication-based validation to a coherence-based model of sufficiency, beyond a procedural adjustment. Because FDA standards likely influence global development strategies, this change will likely extend beyond the United States. Although accelerating access and reducing development costs are legitimate objectives, the reform raises methodological and epistemological concerns. The statistical rationale does not fully incorporate the contextual factors that influence false-positive risk, including prior plausibility and statistical power. Prior plausibility remains informal, and post-marketing evidence seldom restores the epistemic security provided by independent replication. Replication is not merely a statistical redundancy but rather a structural safeguard of robustness and external validity. Without explicit safeguards, increased regulatory flexibility may generate heterogeneity in evidentiary thresholds and progressively weaken the discriminative function of health technology assessment.
BACKGROUND:The systematic conversion of adverse drug reactions (ADRs) into new therapeutic indications represents one of the oldest and most productive sources of repositioned drugs; however, it has not been subjected to formal mechanistic analysis. OBJECTIVES:This study aims to propose a mechanistic taxonomy of convertible ADRs and a structured workflow for translating documented adverse effects into testable repositioning hypotheses. METHODS:In this narrative review, historical cases of ADR-driven repositioning spanning five decades-from sildenafil and minoxidil to semaglutide-were analyzed and classified within a four-dimensional framework organized along the axes of: (1) target identity (on-target versus off-target conversion); (2) dose, route of administration, and reformulation; (3) patient population and clinical context; and (4) pharmacogenomic variability. RESULTS:Each historical case was successfully classified within the proposed taxonomy. A five-step theoretical workflow-comprising signal identification, taxonomic classification, mechanistic verification, feasibility assessment, and preliminary evidence search-was developed to operationalize the framework for prospective hypothesis generation. Computational tools including side-effect similarity networks, network pharmacology, and transcriptomic signature reversal were identified as means to support specific workflow steps. CONCLUSIONS:The proposed taxonomy and workflow provide a replicable, mechanism-informed approach to ADR-driven drug repositioning, while also delineating the pharmacological, regulatory, and methodological challenges that separate a repositioning signal from an approved therapeutic indication.
BACKGROUND:Midazolam (MDZ) is used for sedation in critically ill patients. Although previous studies have examined MDZ pharmacokinetics in ICU patients, the differential effects of acute kidney injury (AKI) and chronic kidney disease (CKD), particularly in cohorts including COVID-19 and non-COVID-19 patients, remain insufficiently characterised. OBJECTIVES:To investigate whether AKI and CKD are associated with dose-normalised exposure of MDZ, 1-hydroxy-midazolam (1-OH-MDZ) and 1-hydroxy-midazolam glucuronide (1-OH-MDZ-G) in critically ill adults. METHODS:In this retrospective observational study, ICU patients receiving continuous intravenous MDZ were included. Plasma concentrations of MDZ, 1-OH-MDZ and 1-OH-MDZ-G were converted to concentration-to-dose (C/D) ratios. Cross-sectional analysis used the first sample per patient; longitudinal analysis included all samples in GEE models. Analyses were adjusted for age, sex, BMI, COVID-19 status, CYP3A4 inhibitor use, APACHE IV score, time since MDZ initiation and CRP. RESULTS:Forty-seven patients contributed 137 samples. In the cross-sectional analysis, C/D ratios of MDZ and 1-OH-MDZ did not differ between patients with and without AKI, whereas 1-OH-MDZ-G C/D ratios were higher in AKI (median [IQR] 14.54 [6.14-26.20] vs. 4.53 [2.52-6.00]; p < 0.01). Longitudinal analysis showed that AKI (β = 0.87), AKI Stage 1 (β = 0.81), AKI Stages 2-3 (β = 0.97) and CKD (β = 0.72) were all significantly associated with higher 1-OH-MDZ-G C/D ratios (all p ≤ 0.03). No significant associations were observed for MDZ or 1-OH-MDZ. CONCLUSION:AKI and CKD are associated with accumulation of 1-OH-MDZ-G in critically ill patients. Monitoring 1-OH-MDZ-G may help identify patients at risk of prolonged sedation and adverse outcomes.
Cardiovascular complications of diabetes mellitus (DM) are responsible for morbidity and mortality in DM patients. Recent evidence highlights the pathophysiological role of the renin-angiotensin-aldosterone system in DM. This study determined whether chronic angiotensin (1-7) (Ang (1-7); 576 μg/kg, s.c., 4 weeks) treatment improves vascular reactivity in mesenteric small resistance arteries of streptozotocin-induced diabetic rats using a wire myograph system and to investigate the underlying mechanisms. Increased sensitivity to vasoconstrictors and decreased responses to endothelium-dependent vasodilators in diabetic rats compared to control rats were partially reversed by Ang (1-7) treatment. Nω-nitro-L-arginine methyl ester (L-NAME, 10-4 M) incubation significantly decreased vasorelaxant responses to acetylcholine (ACH, 10-10-10-5 M) in all groups except the diabetic group. Superoxide dismutase (SOD, 50 U/mL) incubation produced significant increases in vasorelaxant responses to ACH only in the diabetic groups. Glibenclamide (10-5 M), 4-aminopyridine (4-AP, 5 mM), indomethacin (INDO, 10-5 M), or catalase (CAT, 200 U/mL) incubations did not alter ACH responses in any experimental groups, whereas charybdotoxin (ChTX, 10-7 M) or apamin (APA, 10-7 M) produced significant decreases in ACH-induced vasorelaxation, similar in extent in the control and diabetic rats. Following co-incubation with L-NAME and INDO, ACH-induced vasorelaxation in 30 mM KCl pre-contracted arterial rings did not differ significantly between the groups. These findings indicate that improved nitric oxide bioavailability and reduced superoxide activity contribute to the beneficial vascular effects of Ang (1-7) in diabetic rat resistance arteries. Further in vivo studies are warranted to clarify its therapeutic relevance. SUMMARY: Ang (1-7) improves vascular function in diabetic rat resistance arteries. Treatment increases NO bioavailability and reduces oxidative stress. Vasoconstrictor sensitivity is decreased, improving endothelium-dependent vasorelaxation. Ang (1-7) restores vascular responses to endogenous vasoactive agents. Findings suggest Ang (1-7) as a potential target for diabetes-related vascular issues.
BACKGROUND:Cocaine use disorder (CUD) remains a major public health concern, characterized by high relapse rates and the absence of approved pharmacological treatments. Glucagon-like peptide-1 receptor agonists (GLP-1RA), widely prescribed for metabolic disorders, modulate mesolimbic reward pathways and have been proposed as potential modulators of addictive behaviors. OBJECTIVES:To describe clinical observations of GLP-1RA exposure in individuals with CUD identified through the French national vigilance system and to contextualize these findings through a structured narrative review of preclinical and clinical evidence. METHODS:Cases involving GLP-1RA exposure in individuals with ongoing cocaine use were identified through the French national vigilance system and described. A structured literature search (MEDLINE, Embase, Google Scholar) identified original preclinical and clinical studies examining the effects of GLP-1 receptor activation on cocaine-related behavioral and neurobiological outcomes. RESULTS:Two cases involving patient-initiated off-label GLP-1RA use motivated by perceived anti-craving effects were identified through the vigilance system. The literature synthesis included 22 original publications (17 preclinical and 5 human studies). Across experimental models, GLP-1 receptor activation reduced cocaine self-administration, conditioned reward, and relapse-like behaviors. These effects were associated with the attenuation of mesolimbic dopamine signaling, including reduced cocaine-evoked dopamine release and modulation of dopamine transporter function. Human evidence remains limited and heterogeneous. CONCLUSION:GLP-1 receptor signaling represents a biologically plausible target in CUD. However, current clinical evidence remains preliminary. Patient-driven observations underscore the need for controlled clinical trials and continued vigilance monitoring.
This study aimed to evaluate whether TAAR1 (trace amine-associated receptor 1)-associated trace amines (β-phenylethylamine, tryptamine, octopamine, tyramine, and N-methyltyramine) produced cutaneous nociceptive blockade compared to lidocaine. Cutaneous nociceptive blockade was assessed by inhibition of the cutaneous trunci muscle reflex in response to local noxious stimuli. After subcutaneous injection in rats, the nociceptive blockade produced by β-phenylethylamine, tryptamine, octopamine, tyramine, and N-methyltyramine was compared. We demonstrated that, at a dose of 255.3 μmol/kg, β-phenylethylamine, tryptamine, octopamine, tyramine, and N-methyltyramine produced cutaneous nociceptive blockade. At the ED50 (50% effective dose), the relative potency of β-phenylethylamine (85.5 [87.5-93.2] μmol/kg) was lower than that of lidocaine (17.7 [14.6-2.15] μmol/kg; p < 0.01). On an equianesthetic basis (ED25, ED50, and ED75), β-phenylethylamine exhibited a duration of action similar to that of lidocaine. The addition of clonidine or epinephrine did not prolong the duration of action of β-phenylethylamine, whereas the addition of clonidine or epinephrine prolonged the duration of action of lidocaine. Among these five TAAR1-associated trace amines, β-phenylethylamine was the most potent for producing cutaneous nociceptive blockade, tryptamine and octopamine were moderately potent, and tyramine and N-methyltyramine were the least potent. Although β-phenylethylamine was less potent than lidocaine, it exhibited a similar duration of action. Clonidine and epinephrine did not prolong the duration of action of β-phenylethylamine.
Colorectal cancer remains one of the leading causes of cancer-related mortality worldwide, highlighting the need for novel therapeutic agents with multitarget anticancer activity. Probucol is a phenolic compound with well-established antioxidant and anti-inflammatory properties; however, its effects on colorectal cancer cells have not been fully elucidated. The present study aimed to investigate the antitumor effects of probucol on SW480 human colorectal cancer cells and to explore the underlying molecular mechanisms focusing on cell cycle, apoptosis-, and angiogenesis-associated proteins. SW480 cells treated with probucol were subjected to MTT analysis. Cell cycle distribution and apoptosis were evaluated using cell cycle assay, Annexin V binding assay, and multicaspase activity assay. Apoptosis- and angiogenesis-related protein expression was analyzed by western blot, matrix metalloproteinase-9 (MMP-9) mRNA expression was quantified by real-time PCR. Probucol significantly reduced cell viability in a dose-dependent manner and induced cell cycle arrest at the G0/G1 phase. Treatment with probucol markedly increased early and late apoptotic cell populations and enhanced multicaspase activation. The upregulation of pro-apoptotic Bax protein was accompanied by downregulation of antiapoptotic Bcl-2. In addition, probucol significantly suppressed the expression of angiogenesis-associated proteins, including vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS) and also decreased MMP-9 mRNA expression, indicating a potential antimetastatic effect. These findings demonstrate that probucol exerts significant anticancer effects on colorectal cancer cells by inhibiting cell proliferation, inducing apoptosis, and suppressing angiogenesis- and metastasis-related molecular markers. Probucol may represent a promising candidate for further investigation as a potential multitarget agent in colorectal cancer.
PURPOSE:Lower extremity ischemia-reperfusion injury (IRI) triggers systemic inflammation and oxidative stress, causing myocardial remote ischemia-reperfusion injury (MIRI). Current treatments are limited. METHODS:Male Sprague-Dawley rats (n = 6 per group) were assigned to Sham, I/R, I/R + colchicine (C), I/R + HBOT, and I/R + HBOT + C. IRI was induced by 60-min infrarenal aortic clamping and 120-h reperfusion. TREATMENT:Once daily colchicine (0.3 mg/kg) and HBOT (2.5 ATA, 100% O2) twice daily were administered for 5 days post-reperfusion. Cardiac damage, oxidative stress, inflammation, and NRF-2/HO-1 signaling were measured, and electrocardiography (ECG) was performed. RESULTS:I/R caused severe MIRI (Hs-Troponin T: 568.54 pg/mL; heart OSI: 20.98; TNF-α: 582.89 pg/mL). Both monotherapies reduced damage, but HBOT + C was superior: Hs-Troponin T 342.07 pg/mL, OSI 2.26, TNF-α 278.82 pg/mL, and IL-6 12.25 pg/mL, with the upper end of the interquartile range for NRF-2 (41.03) and HO-1 (14.45) activation and restored redox balance (%DIS/NT: 26.31). No differences were observed between groups on the ECG. CONCLUSIONS:Colchicine suppresses inflammation, relieving inhibition of antioxidant defenses, enabling HBOT to fully activate NRF-2/HO-1. This synergistic HBOT + C strategy effectively mitigates MIRI, offering a promising multimodal therapy for remote organ protection after major ischemia.
The effects of obesity on the vasculature and perivascular adipose tissue (PVAT) function vary according to the vascular territory. Nebivolol is a third-generation β-blocker that exerts vasculoprotective effects in obesity. The thoracic and abdominal aorta show different responses under physiological conditions, and their PVATs have a distinctive composition and secretory profile. We hypothesized that obesity would affect thoracic and abdominal aortic responsiveness, as well as their PVAT function, differently and that nebivolol would restore obesity-induced vascular and PVAT changes. To test this hypothesis, male Wistar Hannover rats were fed a hypercaloric diet for 14 weeks. Nebivolol (10 mg/kg/day) was administered via gavage during the last 4 weeks. Obesity increased superoxide (O2•-) generation within the thoracic PVAT, but this response did not result in a loss of its anticontractile effect. Vascular hypercontractility was detected in thoracic aortas, which was mediated by the overproduction of NADPH oxidase-derived O2•- and reduced nitric oxide (NO) bioavailability. Conversely, obesity induced a pro-contractile phenotype in the PVAT of the abdominal aorta. This response occurred in an endothelium-dependent manner, was ameliorated by tiron, and was accompanied by increased O2•- levels. In all cases, nebivolol reversed the effects of obesity. In summary, obesity differentially affects the thoracic and abdominal aorta and their respective PVATs. Nebivolol exerts vasculoprotective effects through antioxidant mechanisms, leading to a reversal of obesity-induced vascular hypercontractility in both aortic segments. The beneficial effects of nebivolol described here could help mitigate cardiovascular complications associated with obesity. Obesity induces PVAT dysfunction. Extending this knowledge, we demonstrated that obesity differentially affects the thoracic and abdominal aorta along with their associated PVATs. This widespread impairment of PVAT function can drive various adverse vascular consequences, including arterial hypertension and aneurysm formation. Critically, peri-aortic PVAT is of particular clinical relevance due to its association with metabolic risk factors and vascular calcification. Nebivolol exerts vasculoprotective effects via antioxidant mechanisms within PVAT, ultimately reversing obesity-induced vascular hypercontractility in both aortic segments. Thus, investigating pharmacological agents that restore PVAT function, as demonstrated here with nebivolol, represents a promising therapeutic strategy to mitigate obesity-related cardiovascular complications.
BACKGROUND:Ketamine is increasingly prescribed in an off-label manner to treat psychiatric disorders, raising concerns about direct-to-consumer advertising and the proliferation of clinics offering ketamine for at-home use without direct medical supervision. OBJECTIVES:We aimed to identify and characterize clinics advertising ketamine for psychiatric conditions online in the New York metropolitan area (New York, New Jersey, and Connecticut), with attention to advertising suggesting ketamine is prescribed for at-home use. METHODS:In 2025, systematic web searches were conducted to identify clinics advertising prescription ketamine for psychiatric indications. Public-facing website content was reviewed to describe clinic characteristics: service delivery modality, clinician credentials, routes of administration, disorders treated, and advertising practices. A generalized linear model was used to delineate correlates of clinics advertising ketamine for at-home use. RESULTS:233 clinics were located; 36.5% prescribed ketamine for at-home use. 51.5% listed a medical doctor as part of their team and 42.9% advertised oral ketamine. Depression was the most commonly listed disorder treated (94.0%) and 21.9% advertised ketamine to treat substance use disorder. In the multivariable model, advertising ketamine for at-home use was more common among clinics advertising oral ketamine (aPR = 4.10, 95% CI: 2.20-7.60) and less common among clinics listing a medical doctor (aPR = 0.54, 95% CI: 0.30-0.99). CONCLUSIONS:Over a third of clinics advertised ketamine for at-home use. A limitation is that we only focused on public-facing websites. Advertising practices and clinician representation suggest clinics may be advertising in a more consumer-oriented manner, underscoring the need for monitoring and clearer guidance to mitigate potential safety risks.
BACKGROUND:There is a complex link between environmental factors, anxiety levels, and ethanol consumption. Social conditions such as loneliness, peer influence, and housing environment shape drinking behaviors and mental health outcomes. The gut microbiota, in the context of the gut-brain axis, may act as a biomarker. OBJECTIVES:This study investigates how different housing conditions impact anxiety-like behavior, ethanol consumption, and gut microbiota in adult male mice. METHODS:Adult male C57BL/6JRj mice were housed under isolated, semi-isolated, or group-housed conditions. Anxiety-like behavior was assessed using the open field (OF), elevated plus maze (EPM), and novelty-suppressed feeding (NSF) tests. Ethanol intake was evaluated using a 4-week intermittent two-bottle choice paradigm. Fecal samples were collected and analyzed by 16S rRNA sequencing to characterize gut microbiota. Behavioral, drinking, and microbiota data were analyzed using ANOVA-based statistics and QIIME2 pipelines. RESULTS:Semi-isolated mice displayed anxiety-like behavior only in the NSF test. Gut microbiota analysis revealed no significant changes in global diversity. Semi-isolated mice exhibited a specific reduction in the Prevotellaceae, particularly Prevotella UCG_001. Housing conditions also influenced ethanol intake: isolated mice exhibited a low initial preference that increased over time, whereas semi-isolated mice showed a high initial preference followed by avoidance. CONCLUSION:Housing conditions strongly shape voluntary ethanol consumption. Semi-isolation was associated with elevated initial alcohol preference and anxiety-like behavior only in the NSF test, while isolation led to a gradual escalation of ethanol intake. These findings highlight the importance of housing environment and anxiety-related factors in alcohol-related preclinical research.
Psychotropic medicines are known to impair cognitive function acutely, but the specific effects of individual substances remain underexplored. This study investigates the effects of dexamphetamine and zolpidem on cognitive performance to quantify the potential risk of intake and their pharmacokinetic and pharmacodynamic relationship. This randomized, double-blind, placebo-controlled trial (EudraCT 2021-005381 - 17) included 60 healthy men aged 26 ± 5 (mean ± SD) years. Participants received a single oral dose of 30-mg dexamphetamine, 5-mg zolpidem, or placebo (n = 20 per group). Cognitive performance was assessed at baseline and 3 and 8 h after dosing using the computerized Psytest system. Dexamphetamine and zolpidem improved sustained attention, with significant reduction of omissions at 8 h. Reaction time improved in both groups, but zolpidem impaired phasic alertness. Working memory remained unchanged. Plasma concentration of dexamphetamine and zolpidem was 70.8 ± 10.4 ng/mL and 39.0 ± 20.7 ng/mL at 3 h and 45.9 ± 7.9 ng/mL and 8.7 ± 6.6 ng/mL at 8 h, respectively. No correlation between drug plasma concentration and cognitive performance measures was demonstrable. Dexamphetamine caused the strongest subjective effects and highest liking ratings, whereas zolpidem elicited greater subjective dislike. Tolerability was best with placebo, followed by zolpidem and dexamphetamine. Single therapeutic doses of dexamphetamine and zolpidem modestly affected cognitive function 3 h after intake, with no relationship between cognitive performance and the study medicines' plasma concentration. Both medicines improve cognition after 8 h but differed in subjective emotional effects. Nevertheless, generalizability is limited by the inclusion of healthy men only. Trial Registration: EudraCT: Nr: 2021-005381-17.
BACKGROUND:Visceral hypersensitivity and impaired intestinal barrier function are hallmark features of irritable bowel syndrome (IBS), linked to activation of corticotropin-releasing factor (CRF), Toll-like receptor 4 (TLR4), and proinflammatory cytokine pathways. Cimetidine, a classical H2 blocker, has been shown to inhibit Na+/H+ exchangers (NHEs), thereby potentially suppressing proinflammatory cytokine release. OBJECTIVES:This study examined whether cimetidine alleviates visceral hypersensitivity and colonic hyperpermeability in rat models of IBS. METHODS:Visceral pain threshold in response to colonic balloon distention was assessed by electromyographic detection of abdominal muscle contractions during colonic balloon distention, and colonic permeability was measured using Evans blue uptake in LPS- and CRF-induced IBS models in male Sprague-Dawley rats. RESULTS:Intragastric cimetidine (20-100 mg/kg daily for 3 days) prevented LPS-induced visceral hypersensitivity and colonic hyperpermeability dose-dependently and also attenuated CRF-induced changes. In contrast, famotidine, another H2 blocker, did not replicate these effects. Amiloride, an NHE inhibitor, mimicked cimetidine's effects, both of which were abolished by intracisternal SB-334867, an orexin 1 receptor antagonist. Furthermore, atropine, sulpiride, and NG-nitro-L-arginine methyl ester, a nitric oxide (NO) synthesis inhibitor, but not scopolamine butylbromide, a peripheral muscarinic receptor antagonist, or domperidone, a peripheral dopamine D2 receptor antagonist, blocked cimetidine's action. CONCLUSION:These findings suggest that cimetidine prevents visceral hypersensitivity and colonic hyperpermeability in IBS models through mechanisms involving central orexin signaling possibly triggered by NHE inhibition, NO, and central muscarinic and dopamine D2 receptor pathways. These findings may provide a basis for future therapeutic approaches targeting IBS.
OBJECTIVE:The objective of this study is to evaluate the influence of alogliptin treatment on the healing process of traumatic oral ulcers. METHODS:Four experimental groups were used: control group (GC) and three test groups treated with oral Alogliptin at 1 (GTA1), 3 (GTA3), and 9 mg/kg/day (GTA9). Ulcer diameter, body weight, glycemic index, colony-forming unit (CFU), and discomfort were analyzed. Histological slides were prepared for healing scores, inflammatory cell counts, collagen deposition analysis, and immunohistochemistry. RESULTS:Alogliptin treatment increased ulcer area (GTA3-7D: 5.2 ± 1.2; GTA9-3D: 11.8 ± 0.8; GTA9-7D: 5.8 ± 1.3; p < 0.001), CFU counts (p = 0.049), and Grimace/discomfort scores (p = 0.02), while reducing body weight gain (p = 0.007) in GTA3 and GTA9 groups. Microscopic analysis revealed higher histopathological scores (p = 0.039), increased mononuclear cells (p = 0.006), reduced polymorphonuclear cells (p < 0.05), and decreased collagen deposition (18.2 ± 2.6; p = 0.031) in GTA9. Lower TLR4 (p = 0.001) and TGF-β (p < 0.001) expression, alongside increased CD31 immunostaining (p < 0.001), were observed in GTA3 and GTA9, as well as reduced TLR2 expression (p = 0.001) in GTA9. CONCLUSION:Alogliptin delays oral ulcer healing by sustaining inflammation, reducing TGF-β expression, and impairing collagen deposition, and may contribute via reduced TLR2/TLR4 expression, increased microbial burden, and decreased TGF-β.
Resistance to B-Raf inhibitors in melanoma poses a significant challenge to effective treatment. This study investigates the synergistic effects of capivasertib, an Akt pathway inhibitor, in combination with B-Raf inhibitors (vemurafenib and encorafenib) in B-Raf-mutated melanoma models. Combination index analysis demonstrated strong synergy between capivasertib and B-Raf inhibitors in melanoma cells. In contrast, combinations of capivasertib with standard chemotherapeutics were antagonistic, underscoring the specificity of the interaction. Mechanistic studies revealed that capivasertib effectively suppressed Akt signaling. Synergism was abolished in Akt-overexpressing cells, confirming Akt's role in the observed combination effects. The combination treatments significantly reduced tumor growth, with tumor volumes in the capivasertib-vemurafenib and capivasertib-encorafenib groups reduced by ~70% relative to control. Notably, this inhibition was sustained for at least 8 weeks, with tumors in the combination groups remaining minimal, while those in the control group reached maximal size by Week 4. Systemic toxicity analyses revealed no significant changes in body weight or serum markers of pancreatic, kidney, or liver function. These findings establish capivasertib and B-Raf inhibitor combinations as a safe and effective strategy for overcoming resistance B-Raf-mutated melanoma, providing new insights into the potential of dual pathway targeting.
Background The application of human hepatic cell lines to early drug discovery and development instead of human primary hepatocytes (HPHs) has been limited because of the low level of drug-metabolizing enzymes (DMEs).Objective The study aimed to evaluate the effects of dexamethasone (DEX) treatment on DME expression, activities, and regulation in HuH-7 hepatoma cells.Methods Expression and transcriptional regulation of major CYPs and UGTs in HuH-7 cells was evaluated by immunoblotting, probe substrate assays, and treatments with nuclear receptor agonists, including DEX, and antagonists.Results DEX increased the expression and activity of cytochrome P450 (CYP) 3A4, uridine 5 '-diphospho-glucuronosyltransferase (UGT) 1A1, and UGT2B7 but had minimal effects on CYP1A2, CYP2B6, or CYP2C9. These augmented activities of CYP3A4, UGT1A1, and UGT2B7 were concentration-dependently inhibited by their corresponding selective inhibitors. DEX-induced upregulation of CYP3A4 protein expression was abolished by co-treatment with the glucocorticoid receptor (GR) inhibitor, but not by co-treatment with the pregnane X receptor (PXR) inhibitor. However, treatment with PXR or constitutive androstane receptor (CAR) agonist did not lead to transcriptional activation of CYP3A4 and 2B6. Only CYP1A1 was transactivated by an aryl hydrocarbon receptor (AhR) ligand.Conclusion Our results suggest that the activities of some major DMEs (CYP3A4, UGT1A1, and UGT2B7) in HuH-7 cells are promoted by DEX treatment possibly through GR activation. However, unlike HPHs, HuH-7 cells fail to show transcriptional regulation of DMEs by PXR or CAR, which limits their suitability for evaluating DME induction potential of investigational drugs.
BACKGROUND:Iohexol clearance (mGFRioh) is a marker of choice for measuring glomerular filtration rate (GFR) in potential living kidney donors. OBJECTIVES:We studied (1) the determinants of mGFRioh variation among kidney donors before and 1 year after donation and (2) the impact of donor mGFRioh change on the gain in estimated GFR 3 months after transplantation in kidney transplant recipients. METHODS:We performed a retrospective, monocentric study in couples of living kidney donors/transplant recipients. In donors, mGFRioh was measured before and 1 year after donation and calculated using the slope-intercept method and the Bröchner-Mortensen correction. In recipients, eGFR was calculated by the CKD-EPI formula before and 3 months after transplantation. Determinants of mGFRioh change in donors and eGFR change in recipients were studied using univariate and multivariate general linear models. RESULTS:Sixty-nine kidney donors (41 women), with a median age of 56 years, were included. The median (10th-90th percentiles) decrease in mGFRioh at 1 year post-donation was 29.7% (13.9-43.0). Determinants of a greater mGFRioh decline (estimate; standard deviation) included female sex (-9.8; 3.3), older age (-0.43; 0.14), and high pre-donation mGFRioh value (-0.24; 0.07). Smoking status and body mass index were not associated with mGFRioh decrease. In kidney recipients, the gain in eGFRCKD-EPI at 3 months post-transplantation was not correlated to mGFRioh variation (before vs. 1 year post-donation) of donors. CONCLUSIONS:Our data identified female sex and age as key determinants of mGFRioh at 1 year post-donation. Donor mGFRioh pre-donation or mGFRioh change 1 year after donation had no impact on early recipient kidney graft function.
Climate change poses the greatest threat to human health in the 21st century. The healthcare sector contributes approximately 5% of global greenhouse gas emissions and has a significant environmental impact. Although clinical trials are crucial for identifying effective and safe treatments and preventing disease, their environmental impact is poorly documented. Our study aimed to assess the environmental impact of a publicly funded, academic clinical trial by adapting life cycle assessment (LCA) methodology to clinical research. We performed a retrospective, simplified, full LCA using the EF 3.0 methodology on a prospective, double-blind, randomised controlled neurosurgery trial. The trial included 202 patients at 18 university hospitals throughout France. To identify hotspots of interest, 16 impact indicators and their combination into a single score were evaluated. The results showed that climate change (or greenhouse gas emissions) was the most important indicator, accounting for almost 30% of the single score. Greenhouse gas emissions were estimated at 31.6 t of carbon dioxide equivalent. The next most important were resource use of fossils (24%), resource use of minerals and metals (12%), and particulate matter emissions (8%). The main hotspots identified were patient transport and travel by clinical research assistants for source data verification. In conclusion, by using a full LCA approach, our study confirms that conducting a clinical trial has a substantial environmental impact, particularly with regard to greenhouse gas emissions. The main hotspots identified were related to patient transport and clinical research assistants' travel. Trial Registration: The SUCRE study (Treatment of Chronic Subdural Hematoma by Corticosteroids: A Prospective Randomised Study)-clinicaltrials.gov identifier: NCT02650609.
Cocaine use has increased both globally and nationally. This trend is accompanied by a rise in clinical complications. The objective of our study was to identify factors observed to be associated with cocaine use disorder, using data from the OPPIDUM program, collecting information directly from patients with substance use disorders recruiting in care or harm reduction facilities. Cocaine users were divided into two groups: cocaine use disorder and simple use. A univariate analysis was performed to compare the groups, followed by a multivariate analysis to identify factors associated with cocaine use disorder. Between 2019 and 2023, 6863 cocaine users (28% of all OPPIDUM participants) from 116 addiction treatment centers were included. Several factors were found to be significantly associated with cocaine use disorder: extreme precariousness (OR = 1.34, 95% CI [1.12-1.61], p = 0.002), cocaine-only use (OR = 1.90, 95% CI [1.45-2.50], p < 0.0001), alcohol dependence (OR = 1.48, 95% CI [1.30-1.68], p < 0.0001), and use of antidepressants and antipsychotics (OR = 2.08, 95% CI [1.37-3.16], p = 0.001; OR = 1.94, 95% CI [1.40-2.69], p < 0.001). These findings highlight several key factors associated with cocaine use disorder and their clinical implications. The study has limitations: potential selection bias, repeated inclusion of the same users across years, multiple modes of cocaine use, and the exploratory nature of the analyses due to uncontrolled alpha risk. Nevertheless, these insights can help healthcare professionals better understand patient profiles and provide more tailored care and prevention strategies.
